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Improved Superoxide Dismutase Activity in Soybean by Low Temperature Polymerase Chain Reaction (PCR)

ZHANG Liyuan1, MA Hongdan1, ZHAO Handan1, XU Dandan1, QU Jing2, GUAN Shuyan1,*, WANG Piwu2,*   

  1. 1. College of Life Sciences, Jilin Agricultural University, Changchun 130118, China;
    2. Faculty of Agronomy, Jilin Agricultural University, Changchun 130118, China
  • Online:2015-03-15 Published:2015-03-17

Abstract:

Objective: This study aims at modifying the activity of superoxide dismutase in soybean by low temperature
PCR. Methods: Six amplified superoxide dismutase genes at low annealing temperatures were inserted into the expression
vector pREP5N to construct the new vectors pPM1, pPM2, pPM3, pPM4, pPM5 and pPM6. These expression vectors
were introduced into E. coli for enzyme expression. The enzyme activity was determined for screening strains with high
enzyme activity. Results: The alignment analysis of cloned sequences versus published MnSOD sequences showed that
the consistency was 86.57% on average. The homology of amino acid sequences was 82.58%. The molecular weights
of the proteins extracted from six strains were approximately 26 kD as determined by SDS-PAGE. Enzyme activity was
significantly improved (P < 0.01) by 1.95 folds on average. Conclusion: The findings of this study provide technical support
for the application of superoxide dismutase.

Key words: superoxide dismutase gene, low temperature polymerase chain reaction, enzyme activity

CLC Number: